Sequential RAS mutations evaluation in cell-free DNA of patients with tissue RAS wild-type metastatic colorectal cancer: the PERSEIDA (Cohort 2) study.

PURPOSE: RAS (KRAS/NRAS) mutational status on a tumor biopsy is mandatory to guide the best treatment in metastatic colorectal cancer (mCRC). Determining the RAS mutational status by tumor-tissue biopsy is essential in guiding the optimal treatment decision for mCRC. RAS mutations are negative predictive factors for the use of EGFR monoclonal antibodies. Cell-free DNA (cfDNA) analysis enables minimally invasive monitoring of tumor evolution. METHODS/PATIENTS: PERSEIDA was an observational, prospective study assessing cfDNA RAS, BRAF and EGFR mutations (using Idylla™) in first-line mCRC, RAS wild-type (baseline tumor-tissue biopsy) patients (cohort 2). Plasma samples were collected before first-line treatment, after 20 ± 2 weeks, and at disease progression. RESULTS: 117 patients were included (103 received panitumumab + chemotherapy as first-line treatment). At baseline, 7 (6.8%) patients had RAS mutations, 4 (3.9%) BRAF mutations and no EGFR mutations were detected (cfDNA, panitumumab + chemotherapy subpopulation [panitumumab + Ch]). The baseline RAS mutational status concordance between tissue and liquid biopsies was 94.0% (93.2%, panitumumab + Ch). At 20 weeks, only one patient in the study (included in the panitumumab + Ch) had an emerging cfDNA RAS mutation. No emerging BRAF or EGFR mutations were reported. At disease progression, 6 patients had emergent mutations not present at baseline (RAS conversion rate: 13.3% [6/45]; 15.0% [6/40], panitumumab + Ch). CONCLUSIONS: The concordance rate between liquid and solid biopsies at baseline was very high, as previously reported, while our results suggest a considerable emergence of RAS mutations during disease progression. Thus, the dynamics of the genomic landscape in ctDNA may provide relevant information for the management of mCRC patients.

Myocarditis in a young male affected with monkeypox infection: a case report.

Background: Concerns on monkeypox as a disease impacting global public health first emerged in May, 2022, and, since that time, has been identified in more than 50 countries. The condition mainly affects men who have sex with other men. Cardiac disease is a rare complication of monkeypox infection. Here, we describe a case of myocarditis in a young male subsequently diagnosed with monkeypox infection. Case summary: A 42-year-old male reported engaging in high-risk sexual behaviours with another male 10 days before presenting to the emergency department with chest pain, fever, maculopapular rash, and a necrotic chin lesion. Electrocardiography revealed diffuse concave ST-segment elevation associated with elevated cardiac biomarkers. Transthoracic echocardiography revealed normal biventricular systolic function without wall motion abnormalities. We excluded other sexually transmitted diseases or viral infections. Cardiac magnetic resonance imaging (MRI) findings suggested myopericarditis involving the lateral wall and adjacent pericardium. The results of polymerase chain reaction (PCR) tests of pharyngeal, urethral, and blood samples were positive for monkeypox. The patient was treated with high-dose non-steroidal anti-inflammatory drugs (NSAIDs) and colchicine and he recovered soon. Discussion: Monkeypox infections are generally self-limited, with most patients experiencing benign clinical outcomes, no hospitalizations, and few complications. This is a rare report of monkeypox complicated with myopericarditis. Management with high-dose NSAIDs and colchicine relieved our patient's symptoms, suggesting a similar clinical outcome as other idiopathic or virus-related myopericarditis.

Evolution of RAS Mutations in Cell-Free DNA of Patients with Tissue RAS Wild-Type Metastatic Colorectal Cancer Receiving First-Line Treatment: The PERSEIDA Study.

The serial analysis of cell-free DNA (cfDNA) enables minimally invasive monitoring of tumor evolution, providing continuous genetic information. PERSEIDA was an observational, prospective study assessing the cfDNA RAS (KRAS/NRAS) mutational status evolution in first-line, metastatic CRC, RAS wild-type (according to baseline tumor tissue biopsy) patients. Plasma samples were collected before first-line treatment, after 20 ± 2 weeks, and at disease progression. One hundred and nineteen patients were included (102 received panitumumab and chemotherapy as first-line treatment-panitumumab subpopulation). Fifteen (12.6%) patients presented baseline cfDNA RAS mutations (n = 14 [13.7%], panitumumab subpopulation) (mutant allele fraction ≥0.02 for all results). No patients presented emergent mutations (cfDNA RAS mutations not present at baseline) at 20 weeks. At disease progression, 11 patients (n = 9; panitumumab subpopulation) presented emergent mutations (RAS conversion rate: 19.0% [11/58]; 17.7% [9/51], panitumumab subpopulation). In contrast, three (5.2%) patients presenting baseline cfDNA RAS mutations were RAS wild-type at disease progression. No significant associations were observed between overall response rate or progression-free survival and cfDNA RAS mutational status in the total panitumumab subpopulation. Although, in patients with left-sided tumors, a significantly longer progression-free survival was observed in cfDNA RAS wild-type patients compared to those presenting cfDNA RAS mutations at any time. Continuous evaluation of RAS mutations may provide valuable insights on tumor molecular dynamics that can help clinical practice.

Cytogenetic evidence supports Avena insularis being closely related to hexaploid oats.

Cytogenetic observations, phylogenetic studies and genome analysis using high-density genetic markers have suggested a tetraploid Avena species carrying the C and D genomes (formerly C and A) to be the donor of all hexaploid oats (AACCDD). However, controversy surrounds which of the three extant CCDD tetraploid species-A. insularis, A. magna and A. murphyi-is most closely related to hexaploid oats. The present work describes a comparative karyotype analysis of these three CCDD tetraploid species and two hexaploid species, A. sativa and A. byzantina. This involved the use of FISH with six simple sequence repeats (SSRs) with the motifs CT, AAC, AAG, ACG, ATC and ACT, two repeated ribosomal sequences, and C genome-specific repetitive DNA. The hybridization pattern of A. insularis with oligonucleotide (AC)10 was also determined and compared with those previously published for A. sativa and A. byzantina. Significant differences in the 5S sites and SSR hybridization patterns of A. murphyi compared to the other CCDD species rule out its being directly involved in the origin of the hexaploids. In contrast, the repetitive and SSR hybridization patterns shown by the D genome chromosomes, and by most of the C genome chromosomes of A. magna and A. insularis, can be equated with the corresponding chromosomes of the hexaploids. Several chromosome hybridization signals seen for A. insularis, but not for A. magna, were shared with the hexaploid oats species, especially with A. byzantina. These diagnostic signals add weight to the idea that the extant A. insularis, or a direct ancestor of it, is the most closely related progenitor of hexaploid oats. The similarity of the chromosome hybridization patterns of the hexaploids and CCDD tetraploids was taken as being indicative of homology. A common chromosome nomenclature for CCDD species based on that of the hexaploid species is proposed.

Delphi consensus on strategies in the management of opioid-induced constipation in cancer patients.

BACKGROUND: Opioid-induced constipation (OIC) is a frequent and bothersome adverse event related with opioid therapy in cancer patients. Despite the high prevalence, medical management of OIC is often uncertain. The current project aimed to investigate expert opinion on OIC management and provide practical recommendations to improve the clinical approach of OIC in cancer patient. METHODS: A modified Delphi method was conducted involving 46 different physicians experts in OIC. Using a structured questionnaire of 67 items this project intended to seek consensus on aspects related to diagnosis, treatment, and quality of life of cancer patients suffering with OIC. RESULTS: After two rounds, a consensus was reached in 91% of the items proposed, all in agreement. Agreement was obtained on OIC definition (95.7%). Objective and patient-reported outcomes included in that definition should be assessed routinely in clinical practice. Responsive to symptom changes and easy-to-use assessment tools were recommended (87.2%). Successful diagnosis of OIC requires increase clinicians awareness of OIC and proactivity to discuss symptoms with their patients (100%). Successful management of OIC requires individualization of the treatment (100%), regular revaluation once is established, and keeping it for the duration of opioid treatment (91.5%). Oral Peripherally Acting µ-Opioid Receptor Agonists (PAMORAs), were considered good alternatives for the treatment of OIC in cancer patients (97.9%). This drugs and laxatives can be co-prescribed if OIC coexist with functional constipation. CONCLUSIONS: The panelists, based on their expert clinical practice, presented a set of recommendations for the management of OIC in cancer patients.

PK-profiling method for identifying the expression of resistance-associated genes in partially resistant oats to crown rust.

BACKGROUND: Protein kinases play a key role in plant cell homeostasis and the activation of defense mechanisms. Partial resistance to fungi in plants is interesting because of its durability. However, the variable number of minor loci associated with this type of resistance hampers the reliable identification of the full range of genes involved. The present work reports the technique of protein kinase (PK)-profiling for the identification of the PK genes induced in the partially resistant oats line MN841801-1 following exposure to the fungus Puccinia coronata. This is the first time this technique has been used with cDNA (complementary DNA) from a suppression subtractive hybridization library obtained after the hybridization of cDNAs from inoculated and mock-inoculated plants. RESULTS: Six degenerate primers based on the conserved domains of protein kinases were used in a PK-profiling assay including cDNA from mock-inoculated leaves and subtracted cDNA. Of the 75.7% of sequences cloned and sequenced that showed significant similarity to resistance genes, 76% were found to code for PKs. Translation and ClustalW2 alignment of each sequence cloned with the complete sequences of the most similar B. distachyon PKs allowed those of the partially resistant oat line to be deduced and characterized. Further, a phylogenetic study carried out after alignment of these B. distachyon PK sequences with the most similar protein sequences of related species also allowed to deduce different functions for the PK cloned. RT-qPCR (Reverse Transcription-quantitative PCR) was analyzed on nine representative sequences to validate the reliability of the employed PK-profiling method as a tool for identifying the expression of resistance-associated genes. CONCLUSIONS: PK-profiling would appear to be a useful tool for the identification of the PKs expressed in oats after challenge by P. coronata, and perhaps other pathogens. Most of the PKs studied are related to receptor-like protein kinases expressed shortly after infection. This is in agreement with previous studies indicating a close relationship between partial resistance and the first layer of defense against pathogen used by plants.

Chromosomal distribution patterns of the (AC)10 microsatellite and other repetitive sequences, and their use in chromosome rearrangement analysis of species of the genus Avena.

Fluorescence in situ hybridization (FISH) was used to determine the physical location of the (AC)10 microsatellite in metaphase chromosomes of six diploid species (AA or CC genomes), two tetraploid species (AACC genome), and five cultivars of two hexaploid species (AACCDD genome) of the genus Avena, a genus in which genomic relationships remain obscure. A preferential distribution of the (AC)10 microsatellite in the pericentromeric and interstitial regions was seen in both the A- and D-genome chromosomes, while in C-genome chromosomes the majority of signals were located in the pericentromeric heterochromatic regions. New large chromosome rearrangements were detected in two polyploid species: an intergenomic translocation involving chromosomes 17AL and 21DS in Avena sativa 'Araceli' and another involving chromosomes 4CL and 21DS in the analyzed cultivars of Avena byzantina. The latter 4CL-21DS intergenomic translocation differentiates clearly between A. sativa and A. byzantina. Searches for common hybridization patterns on the chromosomes of different species revealed chromosome 10A of Avena magna and 21D of hexaploid oats to be very similar in terms of the distribution of 45S and Am1 sequences. This suggests a common origin for these chromosomes and supports a CCDD rather than an AACC genomic designation for this species.

Tyramide Signal Amplification: Fluorescence In Situ Hybridization for Identifying Homoeologous Chromosomes.

Tyramide signal amplification (TSA) fluorescence in situ hybridization (FISH) has been shown as a valuable molecular tool for visualizing specific amplified DNA sequences in chromosome preparations. This chapter describes how to perform TSA-FISH, paying special interest to its two critical steps: probe generation and metaphase plate generation. The potential of physically mapping 12S-globulin sequences by TSA-FISH as a means of identifying homeology among chromosome regions of Avena species was tested and is discussed.

Identification of Genes in a Partially Resistant Genotype of Avena sativa Expressed in Response to Puccinia coronata Infection.

Cultivated oat (Avena sativa), an important crop in many countries, can suffer significant losses through infection by the fungus Puccinia coronata, the causal agent of crown rust disease. Understanding the molecular basis of existing partial resistance to this disease might provide targets of interest for crop improvement programs. A suppressive subtractive hybridization (SSH) library was constructed using cDNA from the partially resistant oat genotype MN841801-1 after inoculation with the pathogen. A total of 929 genes returned a BLASTx hit and were annotated under different GO terms, including 139 genes previously described as participants in mechanisms related to the defense response and signal transduction. Among these were genes involved in pathogen recognition, cell-wall modification, oxidative burst/ROS scavenging, and abscisic acid biosynthesis, as well genes related to inducible defense responses mediated by salicylic and jasmonic acid (although none of which had been previously reported involved in strong responses). These findings support the hypothesis that basal defense mechanisms are the main systems operating in oat partial resistance to P. coronata. When the expression profiles of 20 selected genes were examined at different times following inoculation with the pathogen, the partially resistant genotype was much quicker in mounting a response than a susceptible genotype. Additionally, a number of genes not previously described in oat transcriptomes were identified in this work, increasing our molecular knowledge of this crop.

Susceptibilidad frente a sarampión y varicela en el personal sanitario de un hospital de tercer nivel en Cataluña / Susceptibility to measles and varicella in healthcare workers in a tertiary hospital in Catalonia

Introducción: Estimar la susceptibilidad al sarampión y la varicela del personal sanitario según características demográficas y laborales en un hospital universitario y público de tercer nivel en Cataluña. Métodos Estudio de prevalencia realizado de enero de 2006 a diciembre de 2008 en 2.752 trabajadores a los que se había realizado determinación de serología frente a sarampión y/o varicela mediante test de ELISA en un examen de salud. Se analizaron los datos según sexo, edad, categoría profesional y unidad de trabajo. Resultados Se detectaron 153 profesionales susceptibles a sarampión y 187 a varicela. La susceptibilidad de los trabajadores a sarampión fue del 6,04% (IC 95%: 5,78-6,30) y a varicela del 7,45% (IC 95%: 7,14-7,75). La susceptibilidad a sarampión más elevada fue la de los médicos internos residentes (MIR), con un 14% (IC 95%: 10,8-18,5). En servicios de alto riesgo, que atienden a pacientes especialmente inmunocomprometidos, la susceptibilidad de los trabajadores fue ligeramente superior a la del resto, tanto para sarampión (6,32 vs 5,93%) como para varicela (8,34 vs 7,09%). Los trabajadores nacidos después de 1980 tenían 20 veces (IC 95%: 11,0-37,2) más probabilidad de ser susceptibles a sarampión y 2 veces (IC 95%: 1,2-3,2) más de ser susceptibles a varicela que los nacidos antes de 1965.ConclusiónLa susceptibilidad frente a sarampión en el personal sanitario de nuestro centro es elevada en las cohortes más jóvenes, con cifras superiores a las esperadas en una comunidad con altas coberturas vacunales frente a triple vírica en población pediátrica desde hace muchos años (AU)

Introduction: To estimate the susceptibility to measles and varicella (chickenpox) in healthcare workers in a public tertiary level teaching hospital, in Catalonia. Methods: A prevalence study was conducted from January 2006 to December 2008 on 2,752 workers who had serology performed for the determination of measles or varicella by ELISA test during a health examination. Data were analysed by, sex, age, professional category and work unit. Results: A total of 153 healthcare workers were susceptible to measles and 187 to varicella. The susceptibility of healthcare workers to measles was 6.04% (95% CI: 5.78 to 6.30), and to varicella it was 7.45% (95%CI: 7.14 to 7.75). The highest susceptibility to measles was in resident physicians with 14% (95% CI: 10.8 to18.5). In high-risk services, where highly immunocompromised patients are attended, the susceptibility of workers was slightly higher than the rest to measles (6.32% vs 5.93%) and varicella (8.34% vs 7.09%).Healthcare workers born after 1980 were 20 times (95% CI: 11.0 to 37.2) more likely to be susceptible to measles, and 2 times (95% CI: 1.2 to 3.2) more likely to be susceptible to varicella than those those born before 1965. Conclusions: The susceptibility to measles in healthcare workers in our centre is higher in younger cohorts, with values higher than expected in a community with high vaccination coverage against measles, mumps, rubella vaccine (MMR) in the paediatric population for many years (AU)

[Susceptibility to measles and varicella in healthcare workers in a tertiary hospital in Catalonia]. / Susceptibilidad frente a sarampión y varicela en el personal sanitario de un hospital de tercer nivel en Cataluña.

INTRODUCTION: To estimate the susceptibility to measles and varicella (chickenpox) in healthcare workers in a public tertiary level teaching hospital, in Catalonia. METHODS: A prevalence study was conducted from January 2006 to December 2008 on 2,752 workers who had serology performed for the determination of measles or varicella by ELISA test during a health examination. Data were analysed by, sex, age, professional category and work unit. RESULTS: A total of 153 healthcare workers were susceptible to measles and 187 to varicella. The susceptibility of healthcare workers to measles was 6.04% (95% CI: 5.78 to 6.30), and to varicella it was 7.45% (95% CI: 7.14 to 7.75). The highest susceptibility to measles was in resident physicians with 14% (95% CI: 10.8 to 18.5). In high-risk services, where highly immunocompromised patients are attended, the susceptibility of workers was slightly higher than the rest to measles (6.32% vs 5.93%) and varicella (8.34% vs 7.09%). Healthcare workers born after 1980 were 20 times (95% CI: 11.0 to 37.2) more likely to be susceptible to measles, and 2 times (95% CI: 1.2 to 3.2) more likely to be susceptible to varicella than those those born before 1965. CONCLUSIONS: The susceptibility to measles in healthcare workers in our centre is higher in younger cohorts, with values higher than expected in a community with high vaccination coverage against measles, mumps, rubella vaccine (MMR) in the paediatric population for many years.

Infección por el virus de la inmunodeficiencia humana y geriatría, una nueva realidad / No disponible

Desde 1996, momento en que se generalizó el uso del tratamiento antirretroviral de alta eficacia, se ha producido un cambio pronóstico importante para los pacientes con infección por el virus de la inmunodeficiencia humana, cronificando la infección y presentando expectativas de vida similar, en algunos casos, a la población general no infectada. Por otro lado, los factores de riesgo para la adquisición de la infección están cambiando, siendo actualmente la vía sexual la principal vía de contagio. Los pacientes ancianos han sido considerados tradicionalmente un grupo de bajo riesgo para la infección, pero las mejoras en el tratamiento de la disfunción eréctil y una falsa sensación de seguridad que conlleva a un bajo uso de métodos anticonceptivos de barrera comportan que los pacientes ancianos presenten un riesgo no despreciable para infectarse. El personal sanitario también infraestima el riesgo en estos pacientes, llevando a retrasos diagnósticos y en el inicio del tratamiento, que empeoran el pronóstico de estos pacientes (AU)

Since 1996, with the widespread use of highly active antiretroviral treatment, a shift has important prognostic for patients infected with the human immunodeficiency virus, becoming a chronic infection and presenting life expectancy similar, in some cases, to not infected general population. Moreover, risk factors for acquisition of infection are changing, sexual intercourse is the main risk factor for HIV. Elderly patients have traditionally been considered a low risk group for infection, but improvements in the treatment of erectile dysfunction and a false sense of security that leads to a low use of barrier methods behave that elderly patients present a non-negligible risk to be infected. Health staff also under estimates the risk in these patients, leading to delays in diagnosis and initiation of treatment, which worsen the prognosis of those patients (AU)

Mapping of STS markers obtained from oat resistance gene analog sequences.

Two previously isolated resistance gene analogs (RGAs) of oat have been located as RFLPs in the reference map of Avena byzantina 'Kanota' x Avena sativa 'Ogle' in regions either homologous or homoeologous to loci for resistance to Puccinia coronata, the causal agent of crown rust. In this study, the RGAs were mapped in two recombinant inbred line (RIL) populations that segregate for crown rust resistance: the diploid Avena strigosa x Avena wiestii RIL population (Asw), which has been used for mapping the complex locus PcA, and the hexaploid MN841801-1 x Noble-2 RIL population (MN), in which QTLs have been located. To obtain single-locus markers, RGAs were converted to sequence tagged site (STS) markers using a procedure involving extension of the original RGA sequence lengths by PCR genome walking, amplification and cloning of the parental fragments, and identification of single nucleotide polymorphisms. The procedure successfully obtained STSs from different members of the L7M2 family of sequences, the initial NBS of which have nucleotide similarities of >83%. However, for RGA III2.18, the parental lines were not polymorphic for the STSs assayed. A sequence characterized amplified region (SCAR) marker with features of an RGA had been previously identified for gene Pc94. This marker was also mapped in the above RIL populations. Markers based on RGA L7M2 co-localized with markers defining the QTL Prq1a in linkage group MN3, and were located 15.2 cM from PcA in linkage group AswAC. The SCAR marker for Pc94 was also located in the QTL Prq1a but at 39.5 cM from PcA in AswAC, indicating that the NBS-LRR sequence represented by this marker is not related to PcA. L7M2 was also excluded as a member of the PcA cluster, although it could be an appropriate marker for the Prq1a cluster if chromosome rearrangements are postulated.

Vacunación frente a hepatitis B y trasplantede progenitores hematopoyéticos / Hepatitis B immunization and hematopoietic stem cell transplantation

No disponible